Use of ethynyl alanine amino diol compounds to inhibit enzymatic conversion of angiotensinogen to angiotensin I

ABSTRACT

Compounds characterized generally as ethynyl alanine amino diol derivatives are useful to inhibit enzymatic conversion of angiotensinogen to angiotensin I. Such enzymatic inhibition is useful to treat disorders mediated by plasma renin activity. An example of such disorders is hypertension. Compounds of particular interest are those of Formula I.                    
     wherein A is selected from CO and SO 2  wherein X is selected from oxygen atom and methylene; wherein each of R 1  and R 9  is a group independently selected from hydrido, methyl, ethyl, n-propyl, isopropyl, benzyl, b, b, b-trifluoroethyl, t-butyloxycarbonyl and methoxymethylcarbonyl, and wherein the nitrogen atom to which R 1  and R 9  are attached may be combined with oxygen to form an N-oxide; wherein R 2  is selected from hydrido, methyl, ethyl and isopropyl; wherein R 3  is selected from benzyl, cyclohexylmethyl, phenethyl, imidazolemethyl, pyridylmethyl and 2-pyridylethyl; wherein R 5  is propargyl or a propargyl-containing moiety; wherein R 7  is cyclohexylmethyl; wherein each of R 4  and R 6  is independently selected from hydrido and methyl; wherein R 8  is selected from ethyl, n-propyl, isobutyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclohexylmethyl, cyclohexylethyl, allyl, vinyl and fluorovinyl; wherein each of R 11  and R 12  is independently selected from hydrido, alkyl and phenyl; wherein m is zero; and wherein n is a number selected from zero through three; or a pharmaceutically-acceptable salt thereof.

This is a divisional of application Ser. No. 09/239,493, filed on Jan.28, 1999, now U.S. Pat. No. 6,174,923, which is a continuation ofapplication Ser. No. 09/046,882, filed on Mar. 24, 1998, now U.S. Pat.No. 5,942,548, which is a continuation of application Ser. No.08/783,954 filed on Jan. 16, 1997, now abandoned, which is acontinuation of application Ser. No. 08/586,440 filed on Jan. 16, 1996,now abandoned, which is a continuation of application Ser. No.08/199,211, filed Feb. 23, 1994, now issued as U.S. Pat. No. 5,484,811,which is a continuation of PCT/US92/08842, filed on Oct. 22, 1992, whichis a continuation of Ser. No. 07/783,955 filed on Oct. 29, 1991, nowissued as U.S. Pat. No. 5,227,401 of Gunnar J. Hanson and John S. Baranentitled—ETHYNYL ALANINE AMINO DIOL COMPOUNDS FOR TREATMENT OFHYPERTENSION.

FIELD OF THE INVENTION

Renin-inhibiting compounds are known for control of hypertension. Ofparticular interest herein are compounds useful as renin inhibitingagents.

BACKGROUND OF THE INVENTION

Renin is a proteolytic enzyme produced and secreted into the bloodstreamby the juxtaglomerular cells of the kidney. In the bloodstream, renincleaves a peptide bond in the serum protein angiotensinogen to produce adecapeptide known as angiotensin I. A second enzyme known as angiotensinconverting enzyme, cleaves angiotensin I to produce the octapeptideknown as angiotensin II. Angiotensin II is a potent pressor agentresponsible for vasoconstriction and elevation of cardiovascularpressure. Attempts have been made to control hypertension by blockingthe action of renin or by blocking the formation of angiotensin II inthe body with inhibitors of angiotensin I converting enzyme.

Classes of compounds published as inhibitors of the action of renin onangiotensinogen include renin antibodies, pepstatin and its analogs,phospholipids, angiotensinogen analogs, pro-renin related analogs andpeptide aldehydes.

A peptide isolated from actinomyces has been reported as an inhibitor ofaspartyl proteases such as pepsin, cathepsin D and renin [Umezawa et al,in J. Antibiot. (Tokyo), 23, 259-262 (1970)]. This peptide, known aspepstatin, was found to reduce blood pressure in vivo after theinjection of hog renin into nephrectomized rats [Gross et al, Science,175, 656 (1971)]. Pepstatin has the disadvantages of low solubility andof inhibiting acid proteases in addition to renin. Modified pepstatinshave been synthesized in an attempt to increase the specificity forhuman renin over other physiologically important enzymes. While somedegree of specificity has been achieved, this approach has led to ratherhigh molecular weight hepta- and octapeptides [Boger et al, Nature, 303,81 (1983)]. High molecular weight peptides are generally consideredundesirable as drugs because gastrointestinal absorption is impaired andplasma stability is compromised.

Short peptide aldehydes have been reported as renin inhibitors [Kokubuet al, Biochim. Biophys. Res. Commun., 118, 929 (1984); Castro et al,FEBS Lett., 167, 273 (1984)]. Such compounds have a reactive C-terminalaldehyde group and would likely be unstable in vivo.

Other peptidyl compounds have been described as renin inhibitors. EPAppl. #128,762, published Dec. 18, 1984, describes dipeptide andtripeptide glyco-containing compounds as renin inhibitors [also seeHanson et al, Biochm. Biophys. Res. Comm., 132, 155-161 (1985), 146,959-963 (1987)]. EP Appl. #181,110, published May 14, 1986, describesdipeptide histidine derivatives as renin inhibitors. EP Appl. #186,977published Jul. 9, 1986 describes renin-inhibiting compounds containingan alkynyl moiety, specifically a propargyl glycine moiety, attached tothe main chain between the N-terminus and the C-terminus, such asN-[4(S)-[(N)-[bis(1-naphthylmethyl)acetyl]-DL-propargylglycylamino]-3(S)-hydroxy-6-methylheptanoyl]-L-isoleucinol.EP Appl. #189,203, published Jul. 30, 1986, describespeptidyl-aminodiols as renin inhibitors. EP Appl. #200,406, publishedDec. 10, 1986, describes alkylnaphthylmethylpropionyl-histidylaminohydroxy alkanoates as renin inhibitors. EP Appl. #216,539,published Apr. 1, 1987, describes alkylnaphthylmethylpropionyl aminoacylaminoalkanoate compounds as renin inhibitors orally administered fortreatment of renin-associated hypertension. EP Appl. #229,667, publishedJul. 22, 1987, describes acyl α-aminoacyl aminodiol compounds having apiperazinylcarbonyl or an alkylaminoalkylcarbonyl terminal group at theN-amino acid terminus, such as2(S)-{[(1-piperazinyl)carbonyl]-oxy]-3-phenylpropionyl}-Phe-His amide of2(S)-amino-1-cyclohexyl-3(R), 4(S)-dihydroxy-6-methylheptane. PCTApplication No. WO 87/04349, published Jul. 30, 1987, describesaminocarbonyl aminoacyl hydroxyether derivatives having analkylamino-containing terminal substituent and which are described ashaving renin-inhibiting activity for use in treating hypertension. EPAppl. #300,189 published Jan. 25, 1989 describes amino acid monohydricderivatives having an alkylamino-alkylamino N-terminus and aβ-alanine-histidine or sarcosyl-histidine attached to the main chainbetween the N-terminus and the C-terminus, which derivatives arementioned as useful in treating hypertension. U.S. Pat. No. 4,902,706which issued Feb. 13, 1990 describes a series ofhistidineamide-containing amino alkylaminocarbonyl-H-terminal aminodiolderivatives for use as renin inhibitors. U.S. Pat. No. 5,032,577 whichissued Jul. 16, 1991 describes a series ofhistidineamide-aminodiol-containing renin inhibitors.

DESCRIPTION OF THE INVENTION

Ethynyl alanine amino diol compounds, having utility as renin inhibitorsfor treatment of hypertension in a subject, constitute a family ofcompounds of general Formula I:

wherein A is selected from methylene, CO, SO and SO₂; wherein X isselected from oxygen atom, methylene and NR₁₀ with R₁₀ selected fromhydrido, alkyl and benzyl; wherein each of R₁ and R₉ is a groupindependently selected from hydrido, alkyl, cycloalkyl, alkoxyacyl,haloalkyl, alkoxycarbonyl, benzyloxycarbonyl, loweralkanoyl,haloalkylacyl, phenyl, benzyl, naphthyl, and naphthylmethyl, any one ofwhich groups having a substitutable position may be optionallysubstituted with one or more radicals selected from alkyl, alkoxy,alkenyl, alkynyl, halo, haloalkyl, cyano and phenyl, and wherein thenitrogen atom to which R₁ and R₉ are attached may be combined withoxygen to form an N-oxide; wherein R₂ is selected from hydrido, alkyl,dialkylaminoalkyl, alkylacylaminoalkyl, benzyl and cycloalkyl; whereinR₃ is selected from alkyl, cycloalkylalkyl, acylaminoalkyl, phenylalkyl,naphthylmethyl, aryl, heterocyclicalkyl and heterocycliccycloalkyl,wherein the cyclic portion of any of said phenylalkyl, naphthylmethyl,aryl, heterocyclicalkyl and heterocycliccycloalkyl groups may besubstituted by one or more radicals selected from halo, hydroxy, alkoxyand alkyl; wherein each of R₄ and R₆ is independently selected fromhydrido, alkyl, benzyl and cycloalkyl; wherein R₅ is selected from

wherein V is selected from hydrido, alkyl, cycloalkyl, haloalkyl, benzyland phenyl; wherein each of R₁₃ and R₁₄ is a radical independentlyselected from hydrido, alkyl, alkenyl, alkynyl, cycloalkyl, phenyl,heterocyclic, heterocyclicalkyl and heterocycliccycloalkyl; wherein R₇is selected from substituted or unsubstituted alkyl, cycloalkyl, phenyl,cycloalkylalkyl and phenylalkyl, any one of which may be substitutedwith one or more groups selected from alkyl, hydroxy, alkoxy, halo,haloalkyl, alkenyl, alkynyl and cyano; wherein R₈ is selected fromhydrido, alkyl, haloalkyl, alkylcycloalkyl, cycloalkyl, cycloalkylalkyl,hydroxyalkyl, alkenyl, alkylcycloalkenyl and alkoxycarbonyl; whereineach of R₁₁ and R₁₂ is independently selected from hydrido, alkyl,haloalkyl, dialkylamino and phenyl; and wherein m is zero or one;wherein n is a number selected from zero through five; wherein p is anumber selected from zero through five; and wherein q is a numberselected from zero through five; or a pharmaceutically-acceptable saltthereof.

A preferred family of compounds consists of compounds of Formula Iwherein A is selected from methylene, CO, SO and SO₂; wherein X isselected from oxygen atom, methylene and NR₁₀ with R₁₀ selected fromhydrido, alkyl and benzyl; wherein each of R₁ and R₉ is independentlyselected from hydrido, lower alkyl, haloalkyl, cycloalkyl,alkoxycarbonyl, benzyloxycarbonyl, loweralkanoyl, alkoxyacyl, phenyl andbenzyl, and wherein the nitrogen atom to which R₁ and R₉ are attachedmay be combined with oxygen to form an N-oxide; wherein each of R₂, R₄and R₆ is independently selected from hydrido and alkyl; wherein R₃ isselected from phenylalkyl, naphthylmethyl, cyclohexylalkyl,cyclopentylalkyl, heteroarylalkyl and heteroarylcycloalkyl; wherein R₅is selected from

wherein V is selected from hydrido, alkyl, haloalkyl, benzyl and phenyl;wherein each of R₁₃ and R₁₄ is a radical independently selected fromhydrido, alkyl, alkenyl, alkynyl, cycloalkyl, heteroaryl,heteroarylalkyl and heteroarylcycloalkyl; wherein R₇ is selected fromsubstituted or unsubstituted cyclohexylmethyl and benzyl, either one ofwhich may be substituted with one or more groups selected from alkyl,hydroxy, alkoxy, halo and haloalkyl; wherein R₈ is selected fromhydrido, methyl, ethyl, n-propyl, n-butyl, isobutyl, cycloalkyl,cycloalkylalkyl, hydroxyalkyl, alkenyl, fluoroalkenyl and fluoroalkyl;wherein each of R₁₁ and R₁₂ is independently selected from hydrido,alkyl, dialkylamino and phenyl; wherein m is zero or one; wherein n is anumber selected from zero through five; wherein p is a number selectedfrom zero through five; and wherein q is a number selected from zerothrough five; or a pharmaceutically-acceptable salt thereof.

A more preferred family of compounds consists of compounds of Formula Iwherein A is selected from methylene, CO, SO and SO₂; wherein X isselected from oxygen atom, methylene and NR₁₀ with R₁₀ selected fromhyrido, alkyl and benzyl; wherein each of R₁ and R₉ is independentlyselected from hydrido, alkyl, alkoxyacyl, haloalkyl, alkoxycarbonyl,benzyloxycarbonyl, and benzyl, and wherein the nitrogen atom to which R₁and R₉ are attached may be combined with oxygen to form an N-oxide;wherein each of R₂, R₄ and R₆ is independently selected from hydrido andalkyl; wherein R₃ is selected from benzyl, phenethyl, cyclohexylmethyl,phenpropyl, pyrrolidinyl, piperidinyl, pyrrolidinylmethyl,piperidinylmethyl, pyrazolemethyl, pyrazoleethyl, pyridylmethyl,pyridylethyl, thiazolemethyl, thiazoleethyl, imidazolemethyl,imidazoleethyl, thienylmethyl, thienylethyl, thiazolylcyclopropyl,imidazolecyclopropyl, thienylcyclopropyl, furanylmethyl, furanylethyl,oxazolemethyl, oxazoleethyl, isoxazolemethyl, isoxazoleethyl,pyridazinemethyl, pyridazineethyl, pyrazinemethyl and pyrazineethyl;wherein R₅ is selected from

wherein V is selected from hydrido, alkyl and haloalkyl; wherein each ofR₁₃ and R₁₄ is a radical independently selected from hydrido, alkyl,alkenyl, alkynyl, thiazole and thiazolemethyl; wherein R₇ iscyclohexylmethyl; wherein R₈ is selected from ethyl, n-propyl, isobutyl,cycloalkyl, cycloalkylalkyl, alkenyl, fluoroalkenyl and perfluoropropyl;wherein each of R₁₁ and R₁₂ is independently selected from hydrido,alkyl, dialkylamino and phenyl; wherein m is zero or one; wherein n is anumber selected from zero through five; wherein p is a number selectedfrom zero through five; and wherein q is a number selected from zerothrough five; or a pharmaceutically-acceptable salt thereof.

An even more preferred family of compounds consists of compounds FormulaI wherein A is selected from CO and SO₂; wherein X is selected fromoxygen atom, methylene and NR₁₀ with R₁₀ selected from hydrido andmethyl; wherein each of R₁ and R₉ is independently selected fromhydrido, lower alkyl, alkoxyacyl, alkoxycarbonyl, benzyloxycarbonyl,haloalkyl and benzyl, and wherein the nitrogen atom to which R₁ and R₉are attached may be combined with oxygen to form an N-oxide; wherein R₂is selected from hydrido, methyl, ethyl and isopropyl; wherein R₃ isselected from benzyl, phenethyl, cyclohexylmethyl pyrrolidinyl,piperidinyl, pyrrolidinylmethyl, piperidinylmethyl, pyrazolemethyl,pyrazoleethyl, pyridylmethyl, pyridylethyl, thiazolemethyl,thiazoleethyl, imidazolemethyl, imidazoleethyl, thienylmethyl,thienylethyl, thiazolylcyclopropyl, imidazolecyclopropyl,thienylcyclopropyl, furanylmethyl, furanylethyl, oxazolemethyl,oxazoleethyl, isoxazolemethyl, isoxazoleethyl, pyridazinemethyl,pyridazineethyl, pyrazinemethyl and pyrazineethyl; wherein each of R₄and R₆ is independently selected from hydrido and methyl; wherein R₅ isselected from

wherein V is selected from hydrido, alkyl and trifluoromethyl; whereineach of R₁₃ and R₁₄ is a radical independently selected from hydrido,alkyl and alkynyl; wherein R₇ is cyclohexylmethyl; wherein R₈ isindependently selected from ethyl, n-propyl, isobutyl, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl,cyclobutylmethyl, cyclopentylmethyl, cyclopentylethyl, cyclohexylmethyl,cyclohexylethyl, allyl, vinyl and fluorovinyl; wherein each of R₁₁ andR₁₂ is independently selected from hydrido, alkyl, dialkylamino andphenyl; wherein m is zero; wherein n is a number selected from zerothrough five; wherein p is a number selected from zero through five; andwherein q is a number selected from zero through five; or apharmaceutically-acceptable salt thereof.

A highly preferred family of compounds consists of compounds of FormulaI wherein A is selected from CO and SO₂; wherein X is selected fromoxygen atom and methylene; wherein each of R₁ and R₉ is independentlyselected from hydrido, methyl, ethyl, n-propyl, isopropyl, benzyl,β,β,β-trifluoroethyl, t-butyloxycarbonyl and methoxymethylcarbonyl, andwherein the nitrogen atom to which R₁ and R₉ are attached may becombined with oxygen to form an N-oxide; wherein R₂ is selected fromhydrido, methyl, ethyl and isopropyl; wherein R₃ is selected frombenzyl, cyclohexylmethyl, phenethyl, pyrazolemethyl, pyrazoleethyl,pyridylmethyl, pyridylethyl, thiazolemethyl, thiazoleethyl,imidazolemethyl, imidazoleethyl, thienylmethyl, thienylethyl,furanylmethyl, furanylethyl, oxazolemethyl, oxazoleethyl,isoxazolemethyl, isoxazoleethyl, pyridazinemethyl, pyridazineethyl,pyrazinemethyl and pyrazineethyl; wherein R₅ is selected from

wherein V is selected from hydrido, alkyl and trifluoromethyl; whereineach of R₁₃ and R₁₄ is a radical independently selected from hydrido,methyl, ethyl, propyl and ethynyl; wherein R₇ is cyclohexylmethyl;wherein each of R₄ and R₆ is independently selected from hydrido andmethyl; wherein R₈ is selected from ethyl, n-propyl, isobutyl,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl,cyclohexylmethyl, cyclohexylethyl, fluorovinyl, allyl and vinyl; whereineach of R₁₁ and R₁₂ is independently selected from hydrido, alkyl,dialkylamino and phenyl; wherein m is zero; wherein n is a numberselected from zero through five; wherein p is a number selected fromzero through five; and wherein q is a number selected from zero throughfive; or a pharmaceutically-acceptable salt thereof.

A more highly preferred class of compounds consists of compounds ofFormula I wherein A is selected from CO and SO₂; wherein X is selectedfrom oxygen atom and methylene; wherein each of R₁ and R₉ is a groupindependently selected from hydrido, methyl, ethyl, n-propyl, isopropyl,benzyl, β,β,β-trifluoroethyl, t-butyloxycarbonyl andmethoxymethylcarbonyl, and wherein the nitrogen atom to which R₁ and R₉are attached may be combined with oxygen to form an N-oxide; wherein R₂is selected from hydrido, methyl, ethyl and isopropyl; wherein R₃ isselected from benzyl, cyclohexylmethyl, phenethyl, imidazolemethyl,pyridylmethyl and 2-pyridylethyl; wherein R₅ is selected from

wherein V is selected from hydrido, alkyl and trifluoromethyl; whereineach of R₁₃ and R₁₄ is a radical independently selected from hydrido,methyl and ethynyl; wherein R₇ is cyclohexylmethyl; wherein each of R₄and R₆ is independently selected from hydrido and methyl; wherein R₈ isselected from ethyl, n-propyl, isobutyl, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclohexylmethyl,cyclohexylethyl, allyl, vinyl and fluorovinyl; wherein each of R₁₁ andR₁₂ is independently selected from hydrido, alkyl and phenyl; wherein mis zero; wherein n is a number selected from zero through three; whereinp is a number selected from one through three; and wherein q is zero orone; or a pharmaceutically-acceptable salt thereof.

The term “hydrido” denotes a single hydrogen atom (H). This hydridogroup may be attached, for example, to an oxygen atom to form a hydroxylgroup; or, as another example, one hydrido group may be attached to acarbon atom to form a NR₁₀ group; or, as another example, two hydridogroups may be attached to a carbon atom to form a —CH₂-group. Where theterm “alkyl” is used, either alone or within other terms such as“haloalkyl” and “hydroxyalkyl”, the term “alkyl” embraces linear orbranched radicals having one to about twenty carbon atoms or,preferably, one to about twelve carbon atoms. More preferred alkylradicals are “lower alkyl” radicals having one to about ten carbonatoms. Most preferred are lower alkyl radicals having one to about sixcarbon atoms. The term “cycloalkyl” embraces cyclic radicals havingthree to about ten ring carbon atoms, preferably three to about sixcarbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl andcyclohexyl. The term “haloalkyl” embraces radicals wherein any one ormore of the alkyl carbon atoms is substituted with one or more halogroups, preferably selected from bromo, chloro and fluoro. Specificallyembraced by the term “haloalkyl” are monohaloalkyl, dihaloalkyl andpolyhaloalkyl groups. A monohaloalkyl group, for example, may haveeither a bromo, a chloro, or a fluoro atom within the group. Dihaloalkyland polyhaloalkyl groups may be substituted with two or more of the samehalo groups, or may have a combination of different halo groups. Adihaloalkyl group, for example, may have two fluoro atoms, such asdifluoromethyl and difluorobutyl groups, or two chloro atoms, such as adichloromethyl group, or one fluoro atom and one chloro atom, such as afluoro-chloromethyl group. Examples of a polyhaloalkyl aretrifluoromethyl, 1,1-difluoroethyl, 2,2,2-trifluoroethyl, perfluoroethyland 2,2,3,3-tetrafluoropropyl groups. The term “difluoroalkyl” embracesalkyl groups having two fluoro atoms substituted on any one or two ofthe alkyl group carbon atoms. The terms “alkylol” and “hydroxyalkyl”embrace linear or branched alkyl groups having one to about ten carbonatoms any one of which may be substituted with one or more hydroxylgroups. The term “alkenyl” embraces linear or branched radicals havingtwo to about twenty carbon atoms, preferably three to about ten carbonatoms, and containing at least one carbon-carbon double bond, whichcarbon-carbon double bond may have either cis or trans geometry withinthe alkenyl moiety. The term “alkynyl” embraces linear or branchedradicals having two to about twenty carbon atoms, preferably two toabout ten carbon atoms, and containing at least one carbon-carbon triplebond. The term “cycloalkenyl” embraces cyclic radicals having three toabout ten ring carbon atoms including one or more double bonds involvingadjacent ring carbons. The terms “alkoxy” and “alkoxyalkyl” embracelinear or branched oxy-containing radicals each having alkyl portions ofone to about ten carbon atoms, such as methoxy group. The term“alkoxyalkyl” also embraces alkyl radicals having two or more alkoxygroups attached to the alkyl radical, that is, to form monoalkoxyalkyland dialkoxyalkyl groups. The “alkoxy” or “alkoxyalkyl” radicals may befurther substituted with one or more halo atoms, such as fluoro, chloroor bromo, to provide haloalkoxy or haloalkoxyalkyl groups. The term“alkylthio” embraces radicals containing a linear or branched alkylgroup, of one to about ten carbon atoms attached to a divalent sulfuratom, such as a methythio group. Preferred aryl groups are thoseconsisting of one, two, or three benzene rings. The term “aryl” embracesaromatic radicals such as phenyl, naphthyl and biphenyl. The term“aralkyl” embraces aryl-substituted alkyl radicals such as benzyl,diphenylmethyl, triphenyl methyl, phenyl-ethyl, phenylbutyl anddiphenylethyl. The terms “benzyl” and “phenylmethyl” areinterchangeable. The terms “aryloxy” and “arylthio” denote radicalrespectively, aryl groups having an oxygen or sulfur atom through whichthe radical is attached to a nucleus, examples of which are phenoxy andphenylthio. The terms “sulfinyl” and “sulfonyl”, whether used alone orlinked to other terms, denotes respectively divalent radicals SO andSO₂. The term “aralkoxy”, alone or within another term, embraces an arylgroup attached to an alkoxy group to form, for example, benzyloxy. Theterm “acyl” whether used alone, or within a term such as acyloxy,denotes a radical provided by the residue after removal of hydroxyl froman organic acid, examples of such radical being acetyl and benzoyl.“Lower alkanoyl” is an example of a more prefered sub-class of acyl. Theterm “amido” denotes a radical consisting of nitrogen atom attached to acarbonyl group, which radical may be further substituted in the mannerdescribed herein. The amido radical can be attached to the nucleus of acompound of the invention through the carbonyl moiety or through thenitrogen atom of the amido radical. The term “alkenylalkyl” denotes aradical having a double-bond unsaturation site between two carbons, andwhich radical may consist of only two carbons or may be furthersubstituted with alkyl groups which may optionally contain additionaldouble-bond unsaturation. The term “heterocyclic”, as used alone orwithin groups such as “heterocyclicalkyl”, and “heterocycliccycloalkyl”,(hereinafter referred to as “heterocyclic-containing groups”) embracesradicals having a saturated, or partially unsaturated, or fullysaturated heterocyclic group, wherein the cyclic portion consists of aring system having one ring or two fused rings, which ring systemcontains one, two or three hetero atoms as ring members selected fromnitrogen, oxygen and sulfur, and which ring system has 4 to about 12ring members. Examples of saturated heterocyclic-containing groups arepyrrolidinyl, piperidinyl, pyrrolidinylmethyl, piperidinylmethyl,pyrrolidinylcyclopropyl and piperidinylcyclopropyl. The term“heteroaryl”, whether used alone or within the greater terms“heteroarylalkyl” or “heteroarylcycloalkyl”, denotes a subset of“heterocyclic-containing groups” having a cyclic portion which isfully-unsaturated, that is, aromatic in character, and which has one ortwo hetero atoms as ring members, said hetero atoms selected fromoxygen, sulfur and nitrogen atoms, and which ring system has five or sixring members. The “heteroaryl” ring may be attached to a linear orbranched alkyl radical having one to about ten carbon atoms or may beattached to a cycloalkyl radical having three to about nine carbonatoms. Examples of such heteroarylalkyl or heteroarylcycloalkyl groupsare pyrazolemethyl, pyrazoleethyl, pyridylmethyl, pyridylethyl,thiazolemethyl, thiazoleethyl, imidazolemethyl, imidazoleethyl,thienylmethyl, thienylethyl, furanylmethyl, furanylethyl, oxazolemethyl,oxazoleethyl, thiazolylcyclopropyl, imidazolecyclopropyl,thienylcyclopropyl, isoxazolemethyl, isoxazoleethyl, pyridazinemethyl,pyridazineethyl, pyrazinemethyl and pyrazineethyl. The “heterocyclic”portion or “heteroaryl” portion of the radical, as well as the alkyl orcycloalkyl portion of groups containing a “heterocyclic” or “heteroaryl”portion, may be substituted at a substitutable position with one or moregroups selected from oxo, alkyl, alkoxy, halo, haloalkyl, cyano,aralkyl, aralkoxy, aryl and aryloxy. Such “heterocyclic”,“heterocyclic”-containing group, or “heteroaryl” group may be attachedas a substituent through a carbon atom of the hetero ring system, or maybe attached through a carbon atom of a moiety substituted on a heteroring-member carbon atom, for example, through the methylene substituentof an imidazolemethyl moiety. Also, a heterocyclic orheterocyclic-containing group may be attached through a ring nitrogenatom. For any of the foregoing defined radicals, preferred radicals arethose containing from one to about fifteen carbon atoms.

Specific examples of alkyl groups are methyl, ethyl, n-propyl,isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl,isopentyl, methylbutyl, dimethylbutyl and neopentyl. Typical alkenyl andalkynyl groups may have one unsaturated bond, such as an allyl group, ormay have a plurality of unsaturated bonds, with such plurality of bondseither adjacent, such as allene-type structures, or in conjugation, orseparated by several saturated carbons.

Also included in the family of compounds of Formula I are isomericforms, including diastereoisomers, and the pharmaceutically-acceptablesalts thereof. The term “pharmaceutically-acceptable salts” embracessalts commonly used to form alkali metal salts and to form additionsalts of free acids or free bases. The nature of the salt is notcritical, provided that it is pharmaceutically-acceptable. Suitablepharmaceutically-acceptable acid addition salts of compounds of FormulaI may be prepared from an inorganic acid or from an organic acid.Examples of such inorganic acids are hydrochloric, hydrobromic,hydroiodic, nitric, carbonic, sulfuric and phosphoric acid. Appropriateorganic acids may be selected from aliphatic, cycloaliphatic, aromatic,araliphatic, heterocyclic, carboxylic and sulfonic classes of organicacids, example of which are formic, acetic, propionic, succinic,glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic,glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic,anthranilic, p-hydroxybenzoic, salicyclic, phenylacetic, mandelic,embonic (pamoic), methansulfonic, ethanesulfonic,2-hydroxyethanesulfonic, pantothenic, benzenesulfonic, toluenesulfonic,sulfanilic, mesylic, cyclohexylaminosulfonic, stearic, algenic,β-hydroxybutyric, malonic, galactaric and galacturonic acid. Suitablepharmaceutically-acceptable base addition salts of compounds of FormulaI include metallic salts made from aluminium, calcium, lithium,magnesium, potassium, sodium and zinc or organic salts made fromN,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine,ethylenediamine, meglumine (N-methylglucamine) and procaine. Alsoincluded within the phrase “pharmaceutically-acceptable salts” are“quaternary” salts or salts of “onium” cations, such as ammonium,morpholinium and piperazinium cations, as well as any substitutedderivatives of these cations where the salt is formed on the nitrogenatom lone pair of electrons. All of these salts may be prepared byconventional means from the corresponding compound of Formula I byreacting, for example, the appropriate acid or base with the compound offormula I.

Compounds of Formula I would be useful to inhibit enzymatic conversionof angiotensionogen to angiotensin I. When administered orally, acompound of Formula I would be expected to inhibit plasma renin activityand, consequently, lower blood pressure in a mammalian subject (e.g., ahuman subject). Thus, compounds of Formula I would be therapeuticallyuseful in methods for treating hypertension by administering to ahypertensive subject a therapeutically-effective amount of a compound ofFormula I. The phrase “hypertensive subject” means, in this context, amammalian subject suffering from or afflicted by the effects ofhypertension or susceptible to a hypertensive condition if not treatedto prevent or control such hypertension.

Description of the Synthetic Methods for the Preparation of the ReninInhibitors of the Invention

SYNTHETIC SCHEME 1 Preparation of Compounds of Formula I

A suitably protected amino aldehyde 1 is treated with a Grignard reagentor other organometallic reagent, preferably vinylmagnesium bromide, toobtain the vinyl carbinol 2. This material, suitably protected, isoxidized, preferably with ozone, followed by dimethyl sulfide or zinctreatment, to give intermediate 3. The preceeding process is exemplifiedin Hanson, et al., J. Org. Chem. 50, 5399 (1985). This aldehyde isreacted with an organometallic reagent such as isobutylmagnesiumchloride to give intermediate 4. Other suitable organometallic reagentsinclude ethylmagnesium bromide, vinylmagnesium bromide,cyclopropylmagnesium bromide, and allylmagnesium bromide, but thechoices are not limited to these reagents. After the formation of 4,further transformation of the added side chain is permitted, beforegoing on the next depicted step. For example, the compound 4 derivedfrom the addition of allylmagnesium bromide may be cyclopropanated viadiazomethane and rhodium acetate, to give a cyclopropylmethyl sidechain. Compound 4 is deprotected then coupled, using standardamide/peptide coupling methodology to protected triple bond-containing(ethynyl) amino acid derivatives 5 to give compound 6. These standardcoupling procedures such as the carbodiimide, active ester(N-hydroxysuccinimide), and mixed carbonic anhydride methods are shownin Benoiton, et al. J. Org. Chem. 48, 2939 (1983) and Bodansky, et al.“Peptide Synthesis”, Wiley (1976). Ethynyl-containing amino acidderivatives may be prepared by using procedures such as found inSchollkopf, Tetrahedron 39, 2085 (1983). Intermediate 6 is thendeprotected, then coupled to intermediate 7 using the standardamide/peptide coupling methodology, to give compounds of Formula I.Suitable protecting groups may be selected from among those reviewed byR. Geiger in “The Peptides”, Academic Press, N.Y. vol. 2 (1979). Forexample, P₁ ray by Boc or Cbz; P₂ may be a typical oxygen protectivegroup such as acetyl or t-butyldimethylsilyl.

SYNTHETIC SCHEME 2 Preparation of Compounds of Formula I

Intermediate 7 may be prepared according to the schematic of SyntheticScheme 2. Intermediate 7 is prepared by coupling amine 8 tomono-protected carboxylic acid 9. Carboxylic acid 9 is a mono-activatedmoiety by virtue of a suitable leaving group Q which may be chloride,bromide, fluoride, N-hydroxysuccinimido, p-toluenesulfonyloxy orisobutyloxycarbonyloxy, but is not limited to these groups. Aftercoupling, protecting group P₄ is removed (if P₄ is a benzyl group,hydrogenolysis over palladium-on-carbon (Pd-C) is performed) to giveintermediate amino acid 7.

SYNTHETIC SCHEME 3 Preparation of Compounds of Formula I

Synthetic Scheme 3 describes the preparation of intermediate 8, anon-cyclic diamine. Many of the members of this class, such as ethylenediamine, N,N,N′-trimethylethylene diamine, N,N′-dimethylethylenediamine, N,N′-dimethylpropylene diamine, etc. are commercially availablestarting materials. Other substituted diamines such as compounds 8athrough 8c are obtainable by the procedures depicted in Scheme 3. Forexample, Boc-L-alanine methyl ester is reduced with diisobutylaluminumhydride to give the corresponding aldehyde which is then reductivelyaminated with methylamine, then the Boc group is cleaved to give 8a.Alternatively, the procedure of Miller, et al. J. Med. Chem. 19, 1382(1976) may be employed to give intermediate 8. In another example, asuitably protected diamine is treated with trifluoroacetaldehyde in thepresence of sodium cyanoborohydride to give an trifluoroethylsubstituent on nitrogen, followed by deprotection to give amine 8.

Abbreviations used:

P1 is an N-protecting group; P₂ is H or an oxygen protecting group; P3is an N-protecting group; P4 is an oxygen protecting group such asbenzyl or methyl; Q is a leaving group; Boc is t-butyloxycarbonyl; Cbzis carbobenzoxy.

The following Steps 1-13 constitute specific exemplification of methodsto prepare starting materials and intermediates embraced by theforegoing generic synthetic schemes. Those skilled in the art willreadily understand that known variations of the conditions and processesof the following preparative procedures can be used to prepare thecompounds of Steps 1-13. All temperatures expressed are in degreesCentigrade.

Compounds of Examples 1-3 were prepared by using the proceduresdescribed in the following Steps 1-13:

STEP 1(2R,3S)-N-[(tert-Butyloxy)carbonyl]-3-amino-2-acetoxy-4-phenylbutanal

Ozone/oxygen was bubbled at −70° C. into a solution of(3S,4S)-N-[(tert-Butyloxy)carbonyl]-4-amino-3-acetoxy-5-phenylpentene(2.55 g, 8.0 mmol) (prepared by the method of Hanson, et al., J. Org.Chem. 50, 5399 (1985)) in 100 mL of methylene chloride until a deep bluecolor persisted. Oxygen was introduced until the blue color completelyfaded, then 3.0 mL of Me₂S was added and the solution was allowed towarm to 0-5° C. and stand overnight. The solvent was removed at 0° C.under vacuum yielding the title compound as a thick yellow oil which wasused in the following step without purification.

STEP 2(2S,3R,4S)-N-[(tert-Butyloxy)carbonyl]-2-amino-1-phenyl-3,4-dihydroxy-6-methylheptane

The oil prepared in Step 1 was dissolved under nitrogen in 100 mL of dryTHF and cooled to −70° C. To this solution was added 13 mL (26 mmol) ofa 2.0 M solution of isobutylmagnesium chloride in ether and the stirredmixture was allowed to warm to room temperature and stir for 2 hrs.After decomposition with MeOH/H₂O the mixture was diluted with ether,washed with saturated NH₄Cl solution twice, then dried and the solventsstripped off under vacuum. The residue was allowed to stand overnight in80% MeOH—H₂O containing excess ammonium hydroxide. The MeOH was strippedoff and the mixture was extracted with ether. These extracts werecombined, washed with water, dilute KHSO₄, then dried and evaporated togive 2.36 g of a yellow glass which crystallized from 50 mL of pentaneon standing overnight. The yellow-white powder obtained wasrecrystallized from ether-hexane and furnished the title compound (0.41g) as white, hairy needles, mp 134-136° C., Rf (ether): single spot,0.6. By chromatography of the mother liquors and crystallization of theappropriate fractions, an additional 0.22 g of product, mp 138-139° C.,was obtained.

Anal: Calcd. for C₁₉H₃₁NO₄ (337.45): C, 67.62; H, 9.26; N, 4.15. Found:C, 67.51; H, 9.43; N, 4.24.

STEP 3(2S,3R,4S)-N-[(tert-Butyloxy)carbonyl]-2-amino-1-cyclohexyl-3,4-dihydroxy-6-methylheptane

The diol of Step 2, 0.27 g, was reduced in MeOH with 60 psi H₂ at 60° C.in 3 hrs using 5% Rh/C catalyst. After filtering, the solvent wasstripped off and the white crystals were recrystallized fromCH₂Cl₂-hexane to furnish tiny needles of the title compound, 0.19 g, mp126-128° C.; further recrystallization gave mp 128.5-129.5° C. Rf(ether): single spot, 0.8.

Anal: Calcd. for C₁₉H₃₇NO₄ (343.50): C, 66.43; H, 10.86, N, 4.08. Found:C, 66.43; H, 11.01; N, 4.03.

STEP 4 (2S,3R,4S) 2-Amino-1-cyclohexyl-3, 4-dihydroxy-6-methylheptane

The title compound of Step 3 (10 g) was dissolved 6.9N HCl in dioxane(300 mL). The mixture was stirred for 30 minutes at room temperature.The solvent was removed in vacuo and to the residue was added 5% aqueoussodium hydroxide (30 mL) until a pH of 14 was obtained. This mixture wasextracted with ether and the ether extract was washed with water andbrine, then the solvent was evaporated to give the title compound (7.3g, 100% yield). 300 MHz ¹H NMR: consistent with proposed structure.

Anal. calcd for C₁₄H₂₉NO₂: C, 69.07; H, 12.01; N, 5.78. Found: C, 69.19;H, 12.34; N, 5.78.

STEP 5 D,L Boc-C-propargylglycine

D,L C-propargylglycine (10 g) was suspended in tetrahydrofuran (30 mL).Water (30 mL), potassium carbonate (36.7 g), anddi-tert-butyl-dicarbonate (21.9 g) were added. Additional water wasadded to produce a solution which was stirred for 12 hours at roomtemperature. The organic solvent was then evaporated and the aqueoussolution was washed with ether, then acidified to pH 3 with 1N aqueouscitric acid. The solution was extracted with methylene chloride and thesolvent evaporated to give the title compound (18.9 g, 97% yield), usedwithout further purification.

STEP 6N-[1S,1R*-(Cyclohexylmethyl)-2S*,3R*-dihydroxy-5-methylhexyl]-2-[(1,1-dimethylethoxy)carbonyl]amino]-4-pentynamide

D,L Boc-C-propargylglycine (1.2 g) was dissolved in methylene chloride(5 mL) and N-methyl piperidine (0.57 g) was added. The mixture wascooled to zero degrees centigrade and isobutyl chloroformate (0.78 g)was added. The mixture was stirred for 10 minutes whereupon the titlecompound of Step 4 (1.4 g) in methylene chloride (5 mL) was added andthis mixture stirred for 15 minutes at 0° C. and 4° C. for 12 hours. Thereaction mixture was washed successively with 1N citric acid, saturatedsodium hydrogen carbonate, waster and brine. The organic layer was driedover magnesium sulfate and evaporated to dryness. 300 MHz ¹H NMR:consistent with proposed structure.

STEP 72R*-Amino-N-[1S,1R*-(cyclohexylmethyl)-2S*,3R*-dihydroxy-5-methylhexyl]-4-pentynamide

The title compound of Step 6 (a 1:1 mixture of diastereomers) (0.76 g)was dissolved in a mixture of trifluoroacetic acid (4.9 mL) andmethylene chloride (4.9 mL), and stirred for 30 minutes at roomtemperature. The solvent was then evaporated and the residue taken up inethyl acetate. The organic layer was washed with saturated sodiumhydrogen carbonate, water and brine, then dried over magnesium sulfateand evaporated. The residue was chromatographed on silica gel, elutingwith ethanolchloroform-ammonium hydroxide (15:85:0.5). The fasterrunning of the two diastereomers was collected and evaporated to givethe pure title compound (0.2 g, 34% yield). 300 MHz ¹H NMR: consistentwith proposed structure.

Anal. calcd for C₁₉H₃₄N₂O₃: C, 67.4; H, 10.12; N, 8.31. Found: C, 66.6;H, 10.05; N, 8.02.

STEP 8 2R-Benzyl Butanedioic Acid, 1-Benzyl Ester, DicyclohexylammoniumSalt

To a slurry of 4-(4-methoxybenzyl) itaconate (prepared by the method ofTalley in U.S. Pat. No. 4,939,288) (50 g) in toluene (250 mL) was added1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 30.4 g) in one portion. Then asolution of benzyl bromide (34.2 g) in toluene (50 mL) was addeddropwise over 0.5 hour. The reaction was stirred for 0.5 hour at roomtemperature and then poured into a separatory funnel. The mixture waswashed with 3N HCl, aqueous sodium bicarbonate, brine and dried overmagnesium sulfate. The solvent was evaporated to give a clear mobileliquid (68 g). Chromatography on silica gel, eluting with from 100%hexane to 25% ethyl acetate gave pure 1-(benzyl)-4-(4-methoxybenzyl)itaconate (55 g, 81% yield). A large Fisher-Porter bottle was chargedwith this itaconate (41 g), triethylamine (36 g), palladium acetate(380mg), tri-o-tolylphosphine (1.04 g) and iodobenzene (24.7 g). Thebottle was sealed and flushed with nitrogen and placed in an oil bathand heated for 70 minutes. The residue was chromatographed on silicagel, eluting with 100% hexanes until the less polar impurities wereremoved. Eluting with 10% ethyl acetate in hexane gave the pure phenylitaconate. This compound (23.8 g) was mixed with toluene (200 mL) andthe resulting solution treated with trifluoroacetic acid (30 mL). Thesolution was stirred at room temperature for 1.5 hour and thenevaporated. The residue was taken up in ether (150 mL) and treated withdicyclohexylamine (10.4 g) and stirred at 0° C. whereupon the saltprecipitated. This was isolated by filtration and washed with hexane anddried to give pure 1-benzyl 2-benzylidene succinoatedicyclohexylammonium salt (21.24 g, 78% yield). This benzylidenecompound (20 g) was place in a Fisher-Porter bottle and also added weredegassed methanol (200 mL) and rhodium (R,R)DiPAMP (600 mg) catalyst.The bottle was sealed and flushed with nitrogen then hydrogen. Thereaction was hydrogenated at 40 psig for 15 hours at room temperature.The contents were then poured into a round bottom flask (500 mL) and thesolvent evaporated to give a dark solid. The residue was taken up inboiling isooctane and allowed to stand, with some title compoundcrystallizing (7.34 g). The non-dissolved residue was taken up inboiling dimethoxyethane. This solution was allowed to cool for 12 hours,whereupon crystals of the title compound formed (6.05 g). Combining thetwo crops gave 13.39 g, 66% yield, mp 122-125° C. 300 MHz ¹H NMR:consistent with proposed structure.

STEP 9 2R-Benzyl Butanedioic Acid, 1-Benzyl Ester

The title compound of Step 8 (9.3 g) was suspended in a mixture of water(84 mL) and methanol (8.5 mL). Solid sodium bisulfate (6.12) was addedand the mixture stirred for 5 minutes. The mixture was extracted withmethylene chloride and the combined extracts were dried over magnesiumsulfate and evaporated to dryness. The residue was chromatographed onsilica gel, eluting with methanolchloroform-acetic acid (5:95:0.5), togive the pure title compound (4.3 g, 74% yield).

STEP 10 BenzylαR-[2-[[2-(dimethylamino)ethyl]methylamino]-2-oxoethyl]benzenePropanoate

The title compound of Step 9 (4.3 g) was dissolved in methylene chloride(20 mL) and N-methyl piperidine (1.86 g) was added. The mixture wascooled to 0° C. and isobutylchloroformate (1.97 g) was added. Themixture was stirred for 10 minutes whereupon N,N,N′-trimethylethylenediamine (2.23 g) in methylene chloride (10 mL) was added. This mixturewas stirred at 4° C. for 3 hours, then washed with 1N citric acid,saturated aqueous sodium bicarbonate, water and brine. The solvent wasevaporated to give the title compound (4.7 g, 85% yield). 300 MHz ¹HNMR: consistent with proposed structure.

STEP 11 αR-[2-[[2-(Dimethylamino)ethyl]methylamino]-2-oxoethyl]benzenePropanoic Acid

The title compound of Step 10 (4.6 g) was dissolved in ethanol (50 mL)and hydrogenated over 4% Pd-C at 5 psi at room temperature for 17 hours.The ethanol was evaporated to give the title compound (3 g, 71% yield).300 MHz ¹H NMR: consistent with proposed structure.

STEP 12O-(N-(Dimethylaminoethyl)-N-methyl-aminocarbonyl)-3-L-phenyllactic Acid

Benzyl L-3-phenyllactate (14.28 g) was dissolved in tetrahydrofuran (357mL) and to this was added carbonyl diimidazole (9.78 g) and the mixturewas stirred at room temperature for 4 hours. N,N,N′-trimethylethylenediamine (6.8 g) was added and the mixture stirred for 8 hours. Thesolvent was evaporated and the residue taken up in ether and washed withwater, dried over magnesium sulfate and evaporated to give a yellow oil(13 g, 61% yield); 300 MHz ¹H NMR consistent with proposed structure.This ester was hydrogenated over 4% Pd-C @ 5 psi and room temperaturefor 3.5 hours in tetrahydrofuran. The title compound was obtained as awhite solid (10 g) and recrystallized from methanol.

Anal. calcd for C₁₅H₂₂N2O₄+H₂O: C, 57.68; H, 7.75; N, 8.98. Found: C,57.60; H, 7.82; N, 8.94.

STEP 132R*-Amino-N-[1S,1R*-(cyclohexylmethyl)-2S*,3R*-dihydroxy-5-methylhexyl]-4-pentynamide

The title compound of Step 6 (a 1:1 mixture of diastereomers) (2.3 g)was dissolved in a mixture of trifluoroacetic acid (14 mL) and methylenechloride (14 mL), and stirred for 30 minutes at room temperature. Thesolvent was then evaporated and the residue taken up in ethyl acetate.The organic layer was washed with saturated sodium hydrogen carbonate,water and brine, then dried over magnesium sulfate and evaporated togive the title compound as a mixture of diastereomers (1.8 g, 100%yield). 300 MHz ¹H NMR: consistent with proposed structure.

The following working Examples are provided to illustrate synthesis ofCompounds 1-34 of the present invention and are not intended to limitthe scope thereof. Those skilled in the art will readily understand thatknown variations of the conditions and processes of the followingpreparative procedures can be used to prepare the compounds of theExamples. All temperatures expressed are in degrees Centigrade.

EXAMPLE 1

N1-[1R*-[[[1S,1R*-(cyclohexylmethyl)-2S*,3R*-dihydroxy-5-methylhexyl]amino]carbonyl]-3-butynyl]-N4-[2-(dimethylamino)ethyl]-N4-methyl-2S*-(phenylmethyl)butanediamide

The title compound of Step 11 (110 mg, 0.37 mmol) was dissolved at roomtemperature in a mixture of dimethylformamide (10 mL) and pyridine (132μL) and to this solution was added di-(N-succinimidyl)carbonate (85 mg,0.33 mmol) and dimethylaminopyridine (4 mg). The mixture was stirred for3 hours, whereupon the title amine of Step 7 (103 mg, 0.30 mmol) wasadded, followed by diisopropyl ethylamine (58 μL). This mixture wasallowed to stir for 12 hours. The solvent was then evaporated and theresidue dissolved in ethyl acetate (10 mL). This mixture was washedsuccessively with water and brine, then dried over sodium sulfate andthe solvent evaporated to give crude product (170 mg). The residue waspurified by chromatography on silica gel, eluting withchloroform-ethanol-ammonium hydroxide (84:15:1) to afford the pure titlecompound as a white powder (130 mg, 68% yield) ¹H NMR: 300 MHz spectrumconsistent with proposed structure.

Anal. calcd for C₃₅H₅₆N₄O₅+0.4 H₂O: C, 67.80; H, 9.23; N, 9.04. Found,C, 67.80; H, 9.13; N, 9.02.

EXAMPLE 2

[1R*-[[[1R*-[[[1S,1R*-(Cyclohexylmethyl)-2S*,3R*-dihydroxy-5-methylhexyl]amino]carbonyl]-3-butynyl]amino]carbonyl]-2-phenylethyl)[2-(dimethylamino)ethyl]methylcarbamate

The title acid of Step 12 (220 mg, 0.75 mmol) was coupled to the titleamine of Step 7 (253 mg, 0.75 mmol) using the procedure described forthe preparation of Example 3. The crude product was purified by flashchromatography on silica gel, eluting with chloroform-ethanol-ammoniumhydroxide (84:15:1), to give pure title compound (280 mg, 61% yield). ¹HNMR: 300 MHz spectrum consistent with proposed structure.

Anal. calcd for C₃₄H₅₄N₄ _(O) ₆+1.5H₂O: C, 63.62; H, 8.95; N, 8.73.Found: C, 63.89; H, 8.86; N, 8.28.

EXAMPLE 3

[1R*-[[[1-[[[1S,1R*-(Cyclohexylmethyl)-2S*,3R*-dihydroxy-5-methylhexyl]amino]carbonyl]-3-butynyl]amino]carbonyl]-2-phenylethyl[]2-(dimethylamino)ethyl]methylcarbamate

To a solution of the title compound of Step 12 (128 mg) in methylenechloride (cooled to 0° C.) was added N-methylpiperidine (43 mg),followed by isobutylchloroformate (59 mg). After 5 minutes of stirring,a solution of the title amine of Step 13 (146 mg) in methylene chloridewas added and the resulting mixture stirred for 4 hours. The solutionwas diluted with additional methylene chloride, then washed successivelywith aqueous dilute citric acid, water, aqueous potassium bicarbonate,dried over magnesioum sulfate and the solvent evaporated to dryness togive the title compound (268 mg, 100% yield). 300 MHz ¹H NMR: consistentwith proposed structure.

Anal. calcd for C₃₄H₅₄N₄O₆: C, 66.42; H, 8.85; N, 9.11. Found: C, 65.61;H, 8.91; N, 8.20.

Compounds #4-34, as shown in Table I below, may be synthesized byreference to the foregoing specific and general procedures for preparingcompounds of Formula I.

TABLE I Example Compound No. Structure 4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

BIOLOGICAL EVALUATION

Human Renin Inhibition in Vitro

Compounds of Formula I were evaluated as inhibitors of human renin in anin vitro assay, as follows: This human renin inhibition test has beenpreviously described in detail [Papaioannou et al., Clinical andExperimental Hypertension, A7(9), 1243-1257 (1985)]. Human renin wasobtained from the National Institute for Biological Standards, London.An incubation mixture was prepared containing the following components:in a total volume of 0.25 mL: 100 mM Tris-acetate buffer at pH 7.4,25×10⁻⁶ Goldblatt units of renin, 0.05 mL of plasma from humanvolunteers taking oral contraceptives, 6.0 mM Na-EDTA, 2.4 mMphenylmethyl sulfonyl fluoride, 1.5 mM 8-hydroxyquinoline, 0.4 mg/mLbovine serum albumin (BSA), and 0.024 mg/mL neomycin sulfate. Thismixture was incubated for two hours at 37° C. in the presence or absenceof renin inhibitors. The produced angiotensin I was determined byradioimmunoassay (New England Nuclear kit). Test compounds to be assayedwere dissolved in DMSO and diluted with 100 mM Tris-acetate buffer at pH7.4 containing 0.5% BSA to the appropriate concentration. The finalconcentration of organic solvent in the reaction mixture was less than1%. Control incubations at 37° C. were used to correct for effects oforganic solvent on renin activity.

The in vitro enzymatic conversion of angiotensinogen to angiotensin Iwas inhibited by test compounds of the invention as indicated in TableII, below:

TABLE II Human Renin in vitro Inhibition Data IC₅₀ Human Renin CompoundExample # (nM) Example 1 1 Example 2 4 Example 3 3 Example 4 1

Marmoset Plasma Renin Activity (PRA) Reduction on Oral Administration

The oral activity of renin inhibitor compounds was determined in vivo inMarmoset monkeys using the following procedure. Common Marmoset monkeys(Callithrix jacchus, Charles River, both sexes, body weight 300-400 g)were placed on a modified high-protein low-sodium diet (Purina, St.Louis, Mo.) for 1 week. About 24 hours prior to the administration oftest compound, Lasix (5 mg/kg, im) was given. On the day of the test,the animal was anesthetized with isoflurane, body weight recorded, and abaseline blood sample taken. Then, test compound was givenintragastrically and blood samples were taken in K-EDTA for plasma reninactivity at appropriate time intervals. The PRA was determined by usingthe protocol outlined below. Results are shown below in Table III.Results are expressed in terms of PRA at various time intervals bothbefore and after compound administrations (“Pre” means pretreatmentlevels before compound administrations). Also included is percentinhibition (% INH from pretreatment levels).

PLASMA RENIN ACTIVITY ASSAY

I. Angiotensin I Generation

Plasma Sample 200 ul  PMSF 5% 1 ul Neomycin 10% 3 ul 8-HQ 0.5M 3 ul TES0.5M, pH 7.4 20 ul 

25 ml of the above reaction mixtures, in duplicate, were incubated at37° C. or 0° C. for 2 hours.

II. Determination of Angiotensin I Concentrations

Angiotensin I (AI) concentrations were determined by radioimmunoassayswith a commercial kit from NEN Co.

III. Calculation of Plasma Renin Activity

PRA (ng AI/ml/hr)=(AI at 37°−AI at 0°)/2 hr.

Abbreviation used:

PMSF: Phenylmethylsulonyfluoride

8HQ: 8-Hydroxyquinoline

BSA: Bovine Serum Albumin

TES: N-tris[Hydroxymethyl]methyl-2-aminoethane Sulfonic Acid

EDTA: Ethylenediamine Tetraacetic Acid

TABLE III Effect of Oral Administration of Example 1 Compound onMarmoset PRA (0.1 MPK, IG) Monkey PRA MEAN SC53315 (ngAI/ml/hr) % INH %INH A Pre* 90.1   60 4   95.6 88.7 ± 6 120 5.2 94.2 85.6 ± 7 240 9.289.8  67.5 ± 13 B Pre* 45.8   60 2.8 93.8 120 4.1 91.1 240 14.8  67.8 CPre* 70.2   60 16.4  76.6 120 20   71.5 240 38.7  45   *Time in minutesfrom dosing

Effect of Administration of Example 2 Compound on Marmoset PRA (0.1 MPK,IG)

Monkey PRA MEAN SC53592 (ngAI/ml/hr) % INH % INH A Pre* 118.8   60 33.971.5 81.1 ± 6 120 41.8 64.8 76.6 ± 6 240 49.7 58.2 57.7 ± 0 B Pre* 67.5 60 12.7 81.2 120 14.4 78.7 240 28.5 57.7 C Pre* 42.2  60  3.9 90.7 120 5.7 86.4 240 18.1 57.2 *Time in minutes from dosing

Effect of Oral Administration of Example 3 Compound on Marmoset PRA (0.1 MPK, IG)

Monkey PRA MEAN SC53174 (ngAI/ml/hr) % INH % INH A Pre* 76    60 29.860.7 62.5 ± 2 120 40.3 46.9 54.2 ± 4 240 39.5 48   56.2 ± 5 B Pre* 83.7 60 27.8 66.7 120 33.2 60.3 240 36.1 56.8 C Pre* 105.5   60 42.1 60.1120 47.1 55.3 240 38.1 63.9 *Time in minutes from dosing

Effect of Oral Administration of Example 3 Compound on Marmoset PRA 1.0MPK, IG)

PRA MEAN Monkey (ngAI/ml/hr) % INH % INH A Pre* 185.4  100  60 0  100100 ± 0 120 0  100 100 ± 0 240 0  100 100 ± 0 B Pre* 76.1  60 0  100 1200  100 240 0  100 C Pre* 70.5  60 0  100 120 0  100 *Time in minutesfrom dosing

Determination of Oral Bioavailability

The bioavailability in marmosets and dogs was determined by sampling theblood via the femoral vein at various time points after administeringthe renin inhibitor compounds of the invention in polyethylene glycol400 at an intragastric dose of 10 mg/kg or at an intravenous dose of 1mg/kg. Compound concentration in plasma was determined using thebioassay technique described below. The percent bioavailability wascalculated by dividing the area under the concentration-vs.-time curveobtained from the intragastric experiment by the area under theconcentration-vs.-time curve obtained from the intravenous experiment(adjusting for different doses), and multiplying the result by 100%. Thebioavailibility of Example 1 compound is 27% in marmosets and 33% indogs.

Renin inhibitor plasma concentrations were determined by a bioassay. Theplasma samples were extracted with acetonitrile and the extract wasevaporated to dryness under nitrogen. Residue was dissolved in 4% bovineserum albumin containing 0.9% NaCl and 5% EDTA. The dissolved residue(0.1 ml) was incubated with a reaction mixture containing human plasma(0.12 ml), 5% phenylmethysulfonylfluoride (1.2 ul), 10% neomycin (2.4ul), 0.5 M TES buffer (pH 7.4, 24 ul), and 0.6 mU/ml recombinant humanrenin (1000 U/mg, 50 ul) at 37° C. for 90 minutes. The renin activitywas determined by a standard angiotensin I radioimmunoassay (New EnglandNuclear Corp.). The amount of test compound in the plasma was determinedby comparing the extent of inhibition of renin activity with thatproduced by a known amount of test compound added to plasma and analyzedabove.

Example 1 Compound Oral Bioavailability Studies in Dogs

Estimated Plasma Concentration (ng/ml) Animal Number Time #1 #2 #3 Mean± SE IG at 10 mg/kg: 15 min 105.7  <2.0 265.6 127.7 ± 77.2  30 min 495.9330.4 864.7 563.7 ± 157.9  1 hr 678.2 859.6 859.6 799.1 ± 60.5   2 hr599.2 352.7 554.5 502.1 ± 75.8   4 hr 123.4  68.8 130.3 110.5 ± 20.9   6hr  66.9  47.1  72.4 62.1 ± 7.7   8 hr  33.5  25.4  45.4 34.8 ± 5.8  24hr  6.2  8.4  8.0 7.5 ± 0.7 IV at 1 mg/kg:  2 min 2417.7  3284.4 2982.7  2894.9 ± 254.0   5 min 613.1 680.9 521.1 605.0 ± 46.3  10 min299.0 307.1 218.0 274.7 ± 28.4  15 min 218.0 144.4 146.9 169.8 ± 24.1 30 min 144.7 102.8 122.5 123.3 ± 12.1   1 hr  70.4  67.6  50.0 62.7 ±6.4   2 hr  48.8  56.0  69.8 58.2 ± 6.2   4 hr  18.6  23.5  23.5 21.9 ±1.6   6 hr  10.1  10.2  13.5 11.3 ± 1.1   8 hr  6.4  4.9  6.1 5.8 ± 0.524 hr  2.5  1.5  1.3 1.8 ± 0.4

Summary of Example 1 Oral Bioavailability Studies in Dogs

Animal number #1670841 #1671430 #KOV9 Mean ± SE t1/2 (hr)  5.3  4.4  4.34.7 ± 0.3 Vd (l/kg) 11.5  8.7  8.3 9.5 ± 1.0 Oral (%) Bioavail 35.8 25.638.0 33.1 ± 3.8 

The foregoing data show that the oral bioavailability of Example 1Compound in dogs is 33%, with a terminal half-life of 4.7 hours and avolume of distribution of 9.5 liters/kg.

Example 1 Compound Oral Bioavailability Studies in Marmosets

IG at 10 mg/kg:

Plasma Concentrations (ng/ml)

Test Run A Animal number Time #1 #2 #3 #4 #5 #6 Mean ± SE 30 min — — —9864 7258 9322  1 hr 1340 1989 1751 — — —  2 hr 1561 1525 1811 — — —  3hr — — — 7051 5289 6935  4 hr  742 1766 1217 — — —  5 hr  369 1397  945— — —  6 hr — — — 4914 3177 4412  8 hr — — — 3570 3036 3800

IG at 10 mg/kg:

Plasma Concentrations (ng/ml)

Test Run B Animal number Time #224 #226 #227 #228 #229 #230 Mean ± SE 30min — — — 1344 1016 1126 4988 ± 1749 1 hr 416 5266 12300 — — — 3837 ±1818 2 hr 519 4847 13485 — — — 3958 ± 1997 3 hr — — — 2751 3640 14004511 ± 939  4 hr 459 4502 11470 — — — 3359 ± 1727 5 hr 545 3718  7783 —— — 2460 ± 1174 6 hr — — — 2859 2721 1950 3428 ± 457  8 hr 1805 15661868 2608 ± 400 

IV at 1 mg/kg

Plasma Concentrations (ng/ml)

Test Run C Animal number Mean ± Time #233 #234 #235 #241 #244 #245 SE 2min 13706  21579 16641  — — — 15 min — — — 6756 15283  11628  30 min5858 10359 7921 — — — 1 hr — — — 3091 2665 2793 2 hr  795  2362 1667 — —— 3 hr — — —  486  404  562 4 hr  608  1223 1068 — — — 6 hr — — —  264 208  245

IV at 1 mg/kg

Plasma Concentrations (ng/ml)

Test Run D Animal number Time #233 #234 #235 #237 #239 #240 Mean ± SE 2min 4199 7875 5900 — — — 11650 ± 2772  15 min — — — 7239 2016  897 7303± 2248 30 min 1954 2356 1189 — — — 4939 ± 1514 1 hr — — — 1250 1614 31492427 ± 326  2 hr  554 1004 1055 — — — 1239 ± 271  3 hr — — —  451  5571009 578 ± 90  4 hr  153  587  506 — — — 691 ± 160 6 hr — — —  95  84 158 176 ± 31 

The foregoing data show that the oral bioavailability of Example 1Compound in Marmosets is 27%, with a terminal half-life of 1.5 hours anda volume of distribution of 0.2 liters/kg.

Renin Inhibitor Species Specificity Method

Blood was collected in a 10 ml Becton Dickinson vacutainer tube with 0.1ml of 15% EDTA (K3) solution (15 mg) from normal animals or high reninanimals; i.e., those pre-treated with 5 mg/kg of Lasix® intramuscularly2 times within a 6 hour interval 24 hours prior to bleeding. The bloodwas then centrifuged at 2500 RPM for 20 minutes and the plasma from thevarious species respectively pooled, aliquoted and stored in thefreezer. The human plasma source was a male Caucasian taking aprescription of an angiotensin converting enzyme inhibitor.

The plasma renin activity assay is a modification of the human renininhibition test in that the animal plasma is the source of both therenin substrate and the renin enzyme. In a total volume of 0.1 ml, 90 mMTris-acetate buffer, pH 7.5, 12 mM sodium EDTA, 0.012 mg/ml neomycinsulfate, 0.9 mg/ml bovine serum albumin, 1.61 mM phenylmethyl sulfonylfluoride, 4 mM 8-hydroxyquinoline and 0.09 ml of the animal or humanplasmas were incubated for 2 hours at 37° C. in a shaking water bath orat 4° C. in an ice bath in the presence or absence of renin inhibitors.The produced angiotensin I was determined by radioimmunoassay (NewEngland Nuclear kit).

The amount of angiotensin I generated during the 2 hours at 4° C. wasusually less than 5% of that activity produced at 37° C.; however, the4° C. background values were nevertheless subtracted from the 37° C.ones for the 100% activity values. The renin inhibitors were assayed induplicate using 5 concentrations and the data is expressed as a percentof the 100% total renin activity.

Species Specificity of Example 1 % of Total Renin Activity

Marmoset Cyno Plasma Conc. Human Monkey Monkey Rat 1600 nM  (23-34) 400nM 76% (74-80) 100 nM 93% (91-94)  16 nM 10% 11% 18%  (7-13) (10-12)(11-22)  4 nM 50% 42% 51% (49-51) (38-45) (46-55)  1 nM 90% 75% 81%(80-98) (69-83) (74-89) 0.25 nM  94% (89-98)

The Effect of Example 1 on Marmoset Blood Pressure Reduction onIntravenous Administration

The blood pressure reducing activity of the renin inhibitor compound ofExample 1 was determined in vivo in Marmoset monkeys using the followingprocedure. Common Marmoset monkeys (Callithrix jacchus, Charles River,both sexes, body weight ca. 400 g) were placed on a modified highprotein low sodium diet (Purina, St. Louis, Mo.) for 1 week. 24 hoursprior to the administration of test compound, Lasix (5 mg/kg, im) wasgiven. On the day of the test, the animal was anesthetized withisoflurane, body weight recorded, and the left femoral artery and veinwere catheterized. The animal was allowed to regain consciousness andthe title compound of Example 1 (100 micrograms/kg) was administeredintravenously in 0.1 mL/kg polyethylene glycol 400 at time zero. Meanarterial blood pressure (MABP) in mmHg was then recorded every tenminutes until 120 minutes.

The Effect of Example 1 on Marmoset Blood Pressure Reduction onIntravenous Administration

Time Statistical (minutes) MABP S.E.M. Significance  0 118  6.240 10106  3.210 * 20 97 5.290 30 93 5.290 * 40 92 9.290 * 50 90 9.290 60 893.210 * 70 87 2.310 * 80 88 1.730 * 90 89 4.040 * 100  91 5.290 * 110 93 4.160 120  94 3.610 * *the asterisk denotes statistically significant(p < 0.05) differences from MABP at time zero by paired t-test (n = 3).

These results indicate that Example 1 compound lowers blood pressure insalt-depleted, conscious marmosets.

Also embraced within this invention is a class of pharmaceuticalcompositions comprising one or more compounds of Formula I inassociation with one or more non-toxic, pharmaceutically acceptablecarriers and/or diluents and/or adjuvants (collectively referred toherein as “carrier” materials) and, if desired, other activeingredients. The compounds of the present invention may be administeredby any suitable route, preferably in the form of a pharmaceuticalcomposition adapted to such a route, and in a dose effective for thetreatment intended. Therapeutically effective doses of the compounds ofthe present invention required to prevent or arrest the progress of themedical condition are readily ascertained by one of ordinary skill inthe art. The compounds and composition may, for example, be administeredintravascularly, intraperitoneally, subcutaneously, intramuscularly ortopically.

For oral administration, the pharmaceutical composition may be in theform of, for example, a tablet, capsule, suspension or liquid. Thepharmaceutical composition is preferably made in the form of a dosageunit containing a particular amount of the active ingredient. Examplesof such dosage units are tablets or capsules. These may with advantagecontain an amount of active ingredient from about 1 to 250 mg,preferably from about 25 to 150 mg. A suitable daily dose for a mammalmay vary widely depending on the condition of the patient and otherfactors. However, a dose of from about 0.1 to 3000 mg/kg body weight,particularly from about 1 to 100 mg/kg body weight, may be appropriate.

The active ingredient may also be administered by injection as acomposition wherein, for example, saline, dextrose or water may be usedas a suitable carrier. A suitable daily dose is from about 0.1 to 100mg/kg body weight injected per day in multiple doses depending on thedisease being treated. A preferred daily dose would be from about 1 to30 mg/kg body weight. Compounds indicated for prophylactic therapy willpreferably be administered in a daily dose generally in a range fromabout 0.1 mg to about 100 mg per kilogram of body weight per day. A morepreferred dosage will be a range from about 1 mg to about 100 mg perkilogram of body weight. Most preferred is a dosage in a range fromabout 1 to about 50 mg per kilogram of body weight per day. A suitabledose can be administered, in multiple sub-doses per day. These sub-dosesmay be administered in unit dosage forms. Typically, a dose or sub-dosemay contain from about 1 mg to about 400 mg of active compound per unitdosage form. A more preferred dosage will contain from about 2 mg toabout 200 mg of active compound per unit dosage form. Most preferred isa dosage form containing from about 3 mg to about 100 mg of activecompound per unit dose.

The dosage regimen for treating a disease condition with the compoundsand/or compositions of this invention is selected in accordance with avariety of factors, including the type, age, weight, sex and medicalcondition of the patient, the severity of the disease, the route ofadministration, and the particular compound employed, and thus may varywidely.

For therapeutic purposes, the compounds of this invention are ordinarilycombined with one or more adjuvants appropriate to the indicated routeof administration. If administered per os, the compounds may be admixedwith lactose, sucrose, starch powder, cellulose esters of alkanoicacids, cellulose alkyl esters, talc, stearic acid, magnesium stearate,magnesium oxide, sodium and calcium salts of phosphoric and sulfuricacids, gelatin, acacia gum, sodium alginate, polyvinylpyrrolidone,and/or polyvinyl alcohol, and then tableted or encapsulated forconvenient administration. Such capsules or tablets may contain acontrolled-release formulation as may be provided in a dispersion ofactive compound in hydroxypropylmethyl cellulose. Formulations forparenteral administration may be in the form of aqueous or non-aqueousisotonic sterile injection solutions or suspensions. These solutions andsuspensions may be prepared from sterile powders or granules having oneor more of the carriers or diluents mentioned for use in theformulations for oral administration. The compounds may be dissolved inwater, polyethylene glycol, propylene glycol, ethanol, corn oil,cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride,and/or various buffers. Other adjuvants and modes of administration arewell and widely known in the pharmaceutical art.

Although this invention has been described with respect to specificembodiments, the details of these embodiments are not to be construed aslimitations.

What is claimed is:
 1. A therapeutic method, said method to inhibitenzymatic conversion of angiotensinogen to angiotensin I, comprisingadministering to a patient a therapeutically-effective amount ofcompound of Formula I:

wherein A is selected from methylene, CO, SO and SO₂; wherein X isselected from oxygen atom, methylene and NR₁₀ with R₁₀ selected fromhydrido, alkyl and benzyl; wherein each of R₁ and R₉ is a groupindependently selected from hydrido, alkyl, cycloalkyl, alkoxyacyl,haloalkyl, alkoxycarbonyl, benzyloxycarbonyl, loweralkanoyl,haloalkylacyl, phenyl, benzyl, naphthyl, and naphthylmethyl, any one ofwhich groups having a substitutable position may be optionallysubstituted with one or more radicals selected from alkyl, alkoxy,alkenyl, alkynyl, halo, haloalkyl, cyano and phenyl, and wherein thenitrogen atom to which R1 and R₉ are attached may be combined withoxygen to form an N-oxide; wherein R₂ is selected from hydrido, alkyl,dialkylaminoalkyl, alkylacylaminoalkyl, benzyl and cycloalkyl; whereinR₃ is selected from alkyl, cycloalkylalkyl, acylaminoalkyl, phenylalkyl,naphthylmethyl, aryl, heterocyclicalkyl and heterocycliccycloalkyl,wherein the cyclic portion of any of said phenylalkyl, naphthylmethyl,aryl, heterocyclicalkyl and heterocycliccycloalkyl groups may besubstituted by one or more radicals selected from halo, hydroxy, alkoxyand alkyl; wherein each of R₄ and R₆ is independently selected fromhydrido, alkyl, benzyl and cycloalkyl; wherein R₅ is selected from

wherein V is selected from hydrido, alkyl, cycloalkyl, haloalkyl, benzyland phenyl; wherein each of R₁₃ and R₁₄ is a radical independentlyselected from hydrido, alkyl, alkenyl, alkynyl, cycloalkyl, phenyl,heterocyclic, heterocyclicalkyl and heterocycliccycloalkyl; wherein R₇is selected from substituted or unsubstituted alkyl, cycloalkyl, phenyl,cycloalkylalkyl and phenylalkyl, any one of which may be substitutedwith one or more groups selected from alkyl, hydroxy, alkoxy, halo,haloalkyl, alkenyl, alkynyl and cyano; wherein R₈ is selected fromhydrido, alkyl, haloalkyl, alkylcycloalkyl, cycloalkyl, cycloalkylalkyl,hydroxyalkyl, alkenyl, alkylcycloalkenyl and alkoxycarbonyl; whereineach of R₁₁ and R₁₂ is independently selected from hydrido, alkyl,haloalkyl, dialkylamino and phenyl; and wherein m is zero or one;wherein n is a number selected from zero through five; wherein p is anumber selected from zero through five; and wherein q is a numberselected from zero through five; or a pharmaceutically-acceptable saltthereof.
 2. The method of claim 1 wherein A is selected from methylene,CO, SO and SO₂; wherein X is selected from oxygen atom, methylene andNR₁₀ with R₁₀ selected from hydrido, alkyl and benzyl; wherein each ofR₁ and R₉ is independently selected from hydrido, lower alkyl,haloalkyl, cycloalkyl, alkoxycarbonyl, benzyloxycarbonyl, loweralkanoyl,alkoxyacyl, phenyl and benzyl, and wherein the nitrogen atom to which R₁and R₉ are attached may be combined with oxygen to form an N-oxide;wherein each of R₂, R₄ and R₆ is independently selected from hydrido andalkyl; wherein R₃ is selected from phenylalkyl, naphthylmethyl,cyclohexylalkyl, cyclopentylalkyl, heteroarylalkyl andheteroarylcycloalkyl; wherein R₅ is selected from

wherein V is selected from hydrido, alkyl, haloalkyl, benzyl and phenyl;wherein each of R₁₃ and R₁₄ is a radical independently selected fromhydrido, alkyl, alkenyl, alkynyl, cycloalkyl, heteroaryl,heteroarylalkyl and heteroarylcycloalkyl; wherein R₇ is selected fromsubstituted or unsubstituted cyclohexylmethyl and benzyl, either one ofwhich may be substituted with one or more groups selected from alkyl,hydroxy, alkoxy, halo and haloalkyl; wherein R₈ is selected fromhydrido, methyl, ethyl, n-propyl, n-butyl, isobutyl, cycloalkyl,cycloalkylalkyl, hydroxyalkyl, alkenyl, fluoroalkenyl and fluoroalkyl;wherein each of R₁₁ and R₁₂ is independently selected from hydrido,alkyl, dialkylamino and phenyl; wherein m is zero or one; wherein n is anumber selected from zero through five; wherein p is a number selectedfrom zero through five; and wherein q is a number selected from zerothrough five; or a pharmaceutically-acceptable salt thereof.
 3. Themethod of claim 2 wherein A is selected from methylene, CO, SO and SO₂;wherein X is selected from oxygen atom, methylene and NR₁₀ with R₁₀selected from hydrido, alkyl and benzyl; wherein each of R₁ and R₉ isindependently selected from hydrido, alkyl, alkoxyacyl, haloalkyl,alkoxycarbonyl, benzyloxycarbonyl and benzyl, and wherein the nitrogenatom to which R₁ and R₉ are attached may be combined with oxygen to forman N-oxide; wherein each of R₂, R₄ and R₆ is independently selected fromhydrido and alkyl; wherein R₃ is selected from benzyl, phenethyl,cyclohexylmethyl, phenpropyl, pyrrolidinyl, piperidinyl,pyrrolidinylmethyl, piperidinylmethyl, pyrazolemethyl, pyrazoleethyl,pyridylmethyl, pyridylethyl, thiazolemethyl, thiazoleethyl,imidazolemethyl, imidazoleethyl, thienylmethyl, thienylethyl,thiazolylcyclopropyl, imidazolecyclopropyl, thienylcyclopropyl,furanylmethyl, furanylethyl, oxazolemethyl, oxazoleethyl,isoxazolemethyl, isoxazoleethyl, pyridazinemethyl, pyridazineethyl,pyrazinemethyl and pyrazineethyl; wherein R₅ is selected from

wherein V is selected from hydrido, alkyl and haloalkyl; wherein each ofR₁₃ and R₁₄ is a radical independently selected from hydrido, alkyl,alkenyl, alkynyl, thiazole and thiazolemethyl; wherein R₇ iscyclohexylmethyl; wherein R₈ is selected from ethyl, n-propyl, isobutyl,cycloalkyl, cycloalkylalkyl, alkenyl, fluoroalkenyl and perfluoropropyl;wherein each of R₁₁ and R₁₂ is independently selected from hydrido,alkyl, dialkylamino and phenyl; wherein m is zero or one; wherein n is anumber selected from zero through five; wherein p is a number selectedfrom zero through five; and wherein q is a number selected from zerothrough five; or a pharmaceutically-acceptable salt thereof.
 4. Themethod of claim 3 wherein A is selected from CO and SO₂; wherein X isselected from oxygen atom, methylene and NR₁₀ with R₁₀ selected fromhydrido and methyl; wherein each of R₁ and R₉ is independently selectedfrom hydrido, lower alkyl, alkoxyacyl, alkoxycarbonyl,benzyloxycarbonyl, haloalkyl and benzyl, and wherein the nitrogen atomto which R₁ and R₉ are attached may be combined with oxygen to form anN-oxide; wherein R₂ is selected from hydrido, methyl, ethyl andisopropyl; wherein R₃ is selected from benzyl, phenethyl,cyclohexylmethyl, pyrrolidinyl, piperidinyl, pyrrolidinylmethyl,piperidinylmethyl, pyrazolemethyl, pyrazoleethyl, pyridylmethyl,pyridylethyl, thiazolemethyl, thiazoleethyl, imidazolemethyl,imidazoleethyl, thienylmethyl, thienylethyl, thiazolylcyclopropyl,imidazolecyclopropyl, thienylcyclopropyl, furanylmethyl, furanylethyl,oxazolemethyl, oxazoleethyl, isoxazolemethyl, isoxazoleethyl,pyridazinemethyl, pyridazineethyl, pyrazinemethyl and pyrazineethyl;wherein each of R₄ and R₆ is independently selected from hydrido andmethyl; wherein R₅ is selected from

wherein V is selected from hydrido, alkyl and trifluoromethyl; whereineach of R₁₃ and R₁₄ is a radical independently selected from hydrido,alkyl and alkynyl; wherein R₇ is cyclohexylmethyl; wherein R₈ isindependently selected from ethyl, n-propyl, isobutyl, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl,cyclobutylmethyl, cyclopentylmethyl, cyclopentylethyl, cyclohexylmethyl,cyclohexylethyl, allyl, vinyl and fluorovinyl; wherein each of R₁₁ andR₁₂ is independently selected from hydrido, alkyl, dialkylamino andphenyl; wherein m is zero; wherein n is a number selected from zerothrough five; wherein p is a number selected from zero through five; andwherein q is a number selected from zero through five; or apharmaceutically-acceptable salt thereof.
 5. The method of claim 4wherein A is selected from CO and SO₂; wherein X is selected from oxygenatom and methylene; wherein each of R₁ and R₉ is independently selectedfrom hydrido, methyl, ethyl, n-propyl, isopropyl, benzyl,β,β,β-trifluoroethyl, t-butyloxycarbonyl and methoxymethylcarbonyl, andwherein the nitrogen atom to which R₁ and R₉ are attached may becombined with oxygen to form an N-oxide; wherein R₂ is selected fromhydrido, methyl, ethyl and isopropyl; wherein R₃ is selected frombenzyl, cyclohexylmethyl, phenethyl, pyrazolemethyl, pyrazoleethyl,pyridylmethyl, pyridylethyl, thiazolemethyl, thiazoleethyl,imidazolemethyl, imidazoleethyl, thienylmethyl, thienylethyl,furanylmethyl, furanylethyl, oxazolemethyl, oxazoleethyl,isoxazolemethyl, isoxazoleethyl, pyridazinemethyl, pyridazineethyl,pyrazinemethyl and pyrazineethyl; wherein R₅ is selected from

wherein V is selected from hydrido, alkyl and trifluoromethyl; whereineach of R₁₃ and R₁₄ is a radical independently selected from hydrido,methyl, ethyl, propyl and ethynyl; wherein R₇ is cyclohexylmethyl;wherein each of R₄ and R₆ is independently selected from hydrido andmethyl; wherein R₈ is selected from ethyl, n-propyl, isobutyl,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropylmethyl,cyclohexylmethyl, cyclohexylethyl, fluorovinyl, allyl and vinyl; whereineach of R₁₁ and R₁₂ is independently selected from hydrido, alkyl,dialkylamino and phenyl; wherein m is zero; wherein n is a numberselected from zero through five; wherein p is a number selected fromzero through five; and wherein q is a number selected from zero throughfive; or a pharmaceutically-acceptable salt thereof.
 6. The method ofclaim 5 wherein A is selected from CO and SO₂; wherein X is selectedfrom oxygen atom and methylene; wherein each of R₁ and R₉ is a groupindependently selected from hydrido, methyl, ethyl, n-propyl, isopropyl,benzyl, β,β,β-trifluoroethyl, t-butyloxycarbonyl andmethoxymethylcarbonyl, and wherein the nitrogen atom to which R₁ and R₉are attached may be combined with oxygen to form an N-oxide; wherein R₂is selected from hydrido, methyl, ethyl and isopropyl; wherein R₃ isselected from benzyl, cyclohexylmethyl, phenethyl, imidazolemethyl,pyridylmethyl and 2-pyridylethyl; wherein R₅ is selected from

wherein V is selected from hydrido, alkyl and trifluoromethyl; whereineach of R₁₃ and R₁₄ is a radical independently selected from hydrido,methyl and ethynyl; wherein R₇ is cyclohexylmethyl; wherein each of R₄and R₆ is independently selected from hydrido and methyl; wherein R₈ isselected from ethyl, n-propyl, isobutyl, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cyclopropylmethyl, cyclohexylmethyl,cyclohexylethyl, allyl, vinyl and fluorovinyl; wherein each of R₁₁ andR₁₂ is independently selected from hydrido, alkyl and phenyl; wherein mis zero; wherein n is a number selected from zero through three; whereinp is a number selected from one through three; and wherein q is zero orone; or a pharmaceutically-acceptable salt thereof.
 7. The method ofclaim 6 wherein said compound is selected from compounds, theirtautomers, and the pharmaceutically-acceptable esters and salts thereof,of the group consisting of


8. The method of claim 7 wherein said compound isN1-[1R*-[[[1S,1R*-(cyclohexylmethyl)-2S*,3R*-dihydroxy-5-methylhexyl]amino]carbonyl]-3-butynyl]-N4-[2-(dimethylamino)ethyl]-N4-methyl-2S*-(phenylmethyl)butanediamideor a pharmaceutically-acceptable salt thereof.
 9. The method of claim 7wherein said compound is[1R*-[[[1R*-[[[1S,1R*-(cyclohexylmethyl)-2S*,3R*-dihydroxy-5-methylhexyl]amino]carbonyl]-3-butynyl]amino]carbonyl]-2-phenylethyl)[2-(dimethylamino)ethyl]methylcarbamateor a pharmaceutically-acceptable salt thereof.
 10. The method of claim 7wherein said compound is[1R*-[[[1-[[[1S,1R*-(cyclohexylmethyl)-2S*,3R*-dihydroxy-5-methylhexyl]amino]carbonyl]-3-butynyl)amino]carbonyl]-2-phenylethyl][2-(dimethylamino)ethyl]methylcarbamateor a pharmaceutically-acceptable salt thereof.
 11. The method of claim 7wherein said compound is

or a pharmaceutically-acceptable salt thereof.